Journal: bioRxiv

Article Title: Seminal extracellular vesicles from boar AI doses contain fertility-predictive protein and miRNA cargo and improve sperm physiology

doi: 10.64898/2026.03.16.712050

Figure Lengend Snippet: Cytometry plots and histograms of seminal extracellular vesicles (EVs) characterization. A) Flow cytometry beads of know size (200 nm – 2 µm). B) GFP EVs were used to identify EVs population. C) PBS 1X 0.22 μM filtered. D) Negative control of CellTrace Violet (CTV) staining diluted in PBS. E) High fertility (HF) and reduced fertility (RF) unstained EVs. F) HF CTV-stained EVs. G) RF CTV-stained EVs. H) CTV-stained EVs + human anti CD81-APC antibody (1:50). I) CTV-stained EVs + human anti CD63-PE antibody (1:50). J) CTV-stained EVs + human anti Hsp70-FITC antibody (1:50). K) Lysated EVs with Triton 1X + 0.1% SDS. Photodetectors used: Violet Side Scatter – Height (Violet SSC-H), Blue 525-Area (B525-A), Blue 525-Height (B525-H), Violet 450-Height (V450-H), Blue 585-Height (B585-H) and Red 660-Height (R660-H).

Article Snippet: To immunophenotype the EVs, 25 μL of samples (aproximately 2.0 x 10 9 EVs/mL) were incubated 30 min at 38 °C with conjugated antibodies against specific EVs markers: human anti CD63-PE diluted 1:50 (Ref. 130-118-150, Miltenyi Biotec, Germany), human anti CD81-APC 1:50 (Ref. 130-119-825, Miltenyi Biotec) and human anti Hsp70-FITC 1:50 (Ref. 130-124-700, Miltenyi Biotec).

Techniques: Cytometry, Flow Cytometry, Negative Control, Staining

Journal: Journal of Translational Medicine

Article Title: Eugenoplatin (a phyto-chemo drug conjugate) targets cancer stem cells through inhibition of the β-catenin signaling in triple-negative breast cancer cells

doi: 10.1186/s12967-026-08007-2

Figure Lengend Snippet: Eugenoplatin inhibits the stemness features of breast cancer cells. MDA-MB-231 and BT-20 and SUM149PT cells were either sham-treated (DMSO) or challenged with eugenoplatin (3, 8 and 0.3 µM, respectively) for 24 h. A , cell lysates were prepared and used for immunoblotting analysis, with GAPDH utilized as an internal control. The numbers underneath each band represent fold changes relative to the control (DMSO) after normalization to GAPDH. B , Cells (2 × 10 5 ) were double-stained for both CD24 and CD44, and the proportions of the subpopulation CD44 high /CD24 low were determined by flow cytometry and are shown in the boxes. C , ALDH activity was assessed by the Aldefluor assay and flow cytometry. The numbers in the boxes represent the proportion of ALDH high cells. D , Cells (1 × 10 3 ) were seeded in an ultra-low attachment 96‐well plate containing stem cell‐specific medium (100 µl). The formed spheroids (> 100 μm) were counted. Representative photographs of spheroids (left panels), number of formed spheroids (right panels). Error bars represent mean ± SEM ( n = 3). * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001. E , Cells (1 × 10 3 ) were seeded in an ultra-low attachment 96‐well plate containing stem cell‐specific medium (100 µl). When spheroids were formed, they were either sham-treated (DMSO) or challenged with eugenol or cisplatin using the indicated concentrations for 3 days. Cell viability was determined using the WST1 assay and plotted relative to control cells (DMSO). Error bars represent mean ± SEM ( n = 3). * P ≤ 0.05; *** P ≤ 0.001; $ P ≤ 0.0001. F and G , CD44 high /CD24 low MDA-MB-231cells were sorted by flow cytometry, and then were either sham-treated (DMSO) or challenged with eugenoplatin (3 µM). F , Cell viability was determined using the WST1 assay. Error bars represent mean ± SEM ( n = 3), * P ≤ 0.05. G , The migration ability was determined using the RTCA-DP xCELLigence System

Article Snippet: Cells (3 × 10 5 ) were double-stained with CD44 APC (Miltenyi Biotec) and CD24 PerCP (BioLegend) antibodies for 30 min on ice.

Techniques: Western Blot, Control, Staining, Flow Cytometry, Activity Assay, Migration